文献类型：期刊文献

中文题名：基于PI3K/Akt信号通路探讨地龙蛋白改善糖尿病大鼠勃起功能障碍的作用机制

英文题名：The mechanism of earthworm protein in improving erectile dysfunction in diabetic rats based on PI3K/Akt signaling pathway

作者：刘黎明[1];王新平[2];张爱平[1];剡锐[3];冀小卫[1];王鑫[2];霍耀辉[1];张晓刚[1];邢喜平[2]

第一作者：刘黎明

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃中医药大学附属医院,甘肃兰州730020;[3]兰州大学第一医院,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2023

卷号：37

期号：3

起止页码：26

中文期刊名：中国男科学杂志

外文期刊名：Chinese Journal of Andrology

收录：CSTPCD;;Scopus;CSCD:【CSCD_E2023_2024】；

基金：甘肃省中医药管理局基金项目(GZKP-2021-18);甘肃省中医药防治慢性病重点实验室开放基金(GSMBKY2015-13);甘肃中医药大学附属医院院内课题(gzfy-2019-07);甘肃中医药大学2023年研究生创新创业基金项目;甘肃省自然科学基金项目(23JRRA1198)。

语种：中文

中文关键词：地龙蛋白;糖尿病;勃起功能障碍;细胞凋亡;PI3K/Akt信号通路

外文关键词：earthworm protein;diabetes mellitus;erectile dysfunction;apoptosis;PI3K/Akt signaling pathway

摘要：目的通过检测磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)信号通路相关因子,探讨地龙蛋白对糖尿病性勃起功能障碍(DMED)大鼠的作用机制。方法50只6~8周龄SPF级SD雄鼠通过腹腔注射链脲佐菌素(STZ)构建DMED大鼠模型,同时另取10只6~8周龄SPF级SD雄鼠作为空白组;造模成功后随机分组,西地拉非组给予西地拉非0.005 g/kg灌胃,地龙蛋白低、中、高剂量组分别给予0.045、0.090、0.180 g/kg地龙蛋白灌胃,正常组和模型组均给予生理盐水灌胃,各组10 mL/kg,每日一次。给药4周后,测定大鼠阴茎海绵窦最大内压(ICPmax)和颈动脉压(MAP)以评估大鼠的勃起功能,原位末端标记法(Tunel)检测阴茎海绵体细胞凋亡率,马松(Masson)三色染色观察阴茎海绵体纤维化程度,蛋白免疫印迹法(Western blot)法测定阴茎海绵体中PI3K、Akt、磷酸化蛋白激酶B(pAkt)、半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)表达,实时荧光定量聚合酶链反应法(Real-time PCR)测定上述蛋白mRNA表达。结果与空白组比较,模型组大鼠体质量、阴茎质量、阴茎指数、ICPmax、ICPmax/MAP、阴茎海绵体中PI3K、Akt、pAkt、Bcl-2蛋白及mRNA表达均明显降低(P<0.01),细胞凋亡率、纤维化程度、Caspase-3蛋白及mRNA表达均明显升高(P<0.01)。与模型组比较,各治疗组大鼠体质量、阴茎质量、ICPmax、ICPmax/MAP、阴茎海绵体中PI3K、Akt、pAkt、Bcl-2蛋白及mRNA表达均明显升高(P<0.01),细胞凋亡率、纤维化程度、Caspase-3蛋白及mRNA表达均明显降低(P<0.01)。结论地龙蛋白对DMED大鼠阴茎海绵体有保护作用,在一定程度上改善了阴茎勃起功能,抑制了阴茎海绵体细胞凋亡和纤维化,其机制可能是通过激活PI3K/Akt相关信号通路实现的。
Objective To investigate the underlying mechanism of earthworm protein improving diabetic erectile dysfunction(DMED)by analyzing PI3K/Akt signaling pathway.Methods Fifty 6?8 week?old SD male rats were induced into DMED rat models by intraperitoneal injection of streptozotocin(STZ),and ten 6?8 week?old SD male rats were taken as the normal group;DMED rat models were further divided into four groups.The rats in the sildenafil group was administrated with sildenafil 0.005 g/kg by gavage,and the rats in low,medium and high dose groups with 0.045,0.090,0.180 g/kg of earthworm protein by gavage,and the rats in the normal group and the model group with saline gavage(10 mL/kg),once a day.After 4 weeks intervention,the maximum intracavernosal pressure(ICPmax)and strong arterial pressure(MAP)were measured to assess the erectile function of rats,the apoptosis rate of penile corpus cavernosum cells was detected by in situ end?labeling(Tunel),the fibrosis of penile corpus cavernosum was observed by Masson trichrome staining,the expressions of PI3K,Akt,phosphorylated protein kinase B(pAkt),cysteine aspartate protein hydrolase 3(Caspase?3)and B lymphocyte tumor?2(Bcl?2)were analyzed by Western blot,and the mRNA expressions of these proteins were assayed by real?time fluorescence quantitative polymerase chain reaction(Real?time PCR).Result Compared with those in the normal group,body weight,penile mass,penile index,and ICPmax,ICPmax/MAP,PI3K,Akt,pAkt,Bcl?2 expressions in penile corpus tissues in the model group cavernosum were all significantly decreased(P<0.01),but apoptosis rate,degree of fibrosis,Caspase?3 expression were obviously increased(P<0.01).Compared with those in the model group,the expression of body mass,penile mass,ICPmax,ICPmax/MAP,and P13K,Akt,pAkt,Bcl?2 expressions in the penile corpus cavernosum were all significantly increased in each treatment group(P<0.01),but apoptosis rate,degree of fibrosis,Caspase?3 expression were significantly reduced(P<0.01).Conclusion Earthworm protein had a protective effect on the penile corpus cavernosum of DMED rats by improving erectile function and inhibiting apoptosis and fibrosis of penile corpus cavernosum cells to a certain extent.The mechanism of which may be associated with activation PI3K/Akt signaling pathway.